Target Details

Background: Chemokine(C-C motif) ligand 8, also known as monocyte chemoattractant protein 2(MCP-2), is a protein that in humans is encoded by the CCL8 gene. CCL8 is a small cytokine belonging to the CC chemokine family. The CCL8 protein is produced as a precursor containing 109 amino acids, which is cleaved to produce mature CCL8 containing 75 amino acids. The gene for CCL8 is encoded by 3 exons and is located within a large cluster of CC chemokines on chromosome 17q11.2 in humans. Monocyte chemotactic protein 2(MCP-2) is a CC chemokine that utilizes multiple cellular receptors to attract and activate human leukocytes.

Application Details

Application Details

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Application Notes

Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.

Plate

Pre-coated

Protocol

mouse CCL8 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for CCL8 has been precoated onto 96-well plates. Standards(E.coli, E20-P97) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CCL8 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the mouse CCL8 amount of sample captured in plate.

Assay Procedure

Aliquot 0.1 mL per well of the 1000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, 31.3pg/mL, 15.6pg/mL mouse CCL8 standard solutions into the pre-coated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. We recommend that each mouse CCL8 standard solution and each sample is measured in duplicate.